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Extraction and detection method of marine microalgae fatty acid

A technology of marine microalgae and detection method, which can be applied to measurement devices, instruments, scientific instruments, etc., and can solve the problems of inefficiency, inconvenience, and inaccurate detection results.

Pending Publication Date: 2021-03-23
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention provides a method for extracting and detecting marine microalgae fatty acids in order to overcome the problems of inefficiency, inconvenience, and inaccurate detection results in the extraction and detection method of microalgae fatty acids in the prior art

Method used

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  • Extraction and detection method of marine microalgae fatty acid
  • Extraction and detection method of marine microalgae fatty acid
  • Extraction and detection method of marine microalgae fatty acid

Examples

Experimental program
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Effect test

Embodiment 1

[0044] A method for extracting and detecting marine microalgae fatty acids, comprising the following steps:

[0045] (1) Collection and crushing of Heterocurvium akashiwo

[0046] Take 240 ml of Heterosigma akashiwo based on the end of logarithmic growth, freeze the sample quickly with liquid nitrogen (to inhibit lipase activity), thaw and collect the cells by centrifugation (6000rpm, 4°C), the fresh weight of the cells is 15- 75 mg, and sonicated for 30 minutes in a cell disruptor, took 2 mL of chloroform:methanol (2:1, v / v) to extract lipids, and centrifuged at 12,000 rpm for 10 minutes at room temperature. The hydrophobic phase was extracted as a lipid extract, which was added with 0.01% (w / w) butylated hydroxytoluene (BHT) as an antioxidant at -20°C.

[0047] (2) Separation of neutral and polar lipids

[0048] After the extract was evaporated to dryness and washed three times with 1 mL of chloroform:methanol (98:2, v / v) to recover, the lipid extract was deposited on a so...

Embodiment 2

[0062] The difference from Example 1 is that the specific parameters of step (1), step (2), and step (3) are set, specifically in step (1), the collection and crushing of microalgae; Algae, freeze the sample in liquid nitrogen, centrifuge the sample after thawing, collect the cells after centrifugation, and then use a cell disruptor to sonicate the cells, take chloroform and methanol with a volume ratio of 3:2 to extract lipids, and then After centrifugation at room temperature, the hydrophobic phase was extracted as a lipid extract, which was added with 0.01% (w / w) butylated hydroxytoluene (BHT) as an antioxidant at -20°C.

[0063] In step (2) to separate neutral and polar lipids, evaporate the lipid extract to dryness and recover after washing with chloroform and methanol with a volume ratio of 95:5, deposit the lipid extract on a solid-phase extraction column filled with silica gel Above, neutral lipids were eluted with chloroform:methanol at a volume ratio of 95:5, followe...

Embodiment 3

[0066] The difference from Example 1 lies in the specific parameter settings of step (1), step (2) and step (3), specifically in step (1), the collection and crushing of microalgae; Algae, freeze the sample in liquid nitrogen, centrifuge the sample after thawing, collect the cells after centrifugation, and then use a cell disruptor to sonicate the cells, take chloroform and methanol with a volume ratio of 5:3 to extract lipids, and then After centrifugation at room temperature, the hydrophobic phase was extracted as a lipid extract, which was added with 0.01% (w / w) butylated hydroxytoluene (BHT) as an antioxidant at -20°C.

[0067] In step (2), neutral and polar lipids are separated, the lipid extract is evaporated to dryness and recovered after washing with chloroform and methanol at a volume ratio of 96:4, and the lipid extract is deposited on a solid phase filled with silica gel On the extraction column, neutral lipids were eluted with chloroform and methanol at a volume ra...

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Abstract

The invention relates to the technical field of extraction and detection of microalgae cells, in particular to an extraction and detection method of marine microalgae fatty acid. The extraction and detection method of marine microalgae fatty acid comprises the following steps of (1) collecting and crushing microalgae, (2) separating neutral and polar lipids, (3) lipid fatty acid analysis, and (4)GC-MS analysis. The method disclosed by the invention can be used for efficiently, conveniently and accurately extracting and detecting the microalgae fatty acid, and is a key step for researching fatty acid composition and metabolic pathways thereof.

Description

technical field [0001] The present invention relates to the technical field of extraction and detection of microalgae cells, more specifically, to a method for extraction and detection of marine microalgae fatty acids Background technique [0002] The prevention and resource utilization of harmful red tide algae is an important field of marine ecological governance and sustainable development. Lipids are abundant in microalgae, which produce many different kinds of lipids, such as triglycerides and diglycerides, phospholipids and glycolipids, hydrocarbons, etc. Among them, the phospholipids in the membrane structure of algae occupy the main part of the lipid content. In the field of harmful algae management, the change of the membrane structure composition of microalgae has been used as an important indicator to measure the physiological response of microalgae to external stress. ; It has been reported that the algal toxins produced by algae are mainly membrane lipids. Ther...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/14
CPCG01N30/02G01N30/06G01N30/14
Inventor 叶锦韶赵玲林泽宏潘国强
Owner JINAN UNIVERSITY
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